BK generation from HK and LK. For all reactions, samples were collected at the indicated time points, and BK concentration was determined by ELISA. (A) Plasma-derived HK (200 nM) incubated with 2 nM of PKa (orange), and plasma-derived LK (200 nM) incubated with 50 (light green) or 2 nM of PKa (blue). (B) Plasma-derived HK (200 nM) incubated with 2 nM of PKa (orange), 40 nM of Plm-Glu³¹¹ (red), or 40 nM of Plm-Lys³¹¹ (blue). (C) Plasma-derived LK (200 nM) incubated with 50 nM of PKa (orange), 40 nM of Plm-Glu³¹¹ (red), or 40 nM of Plm-Lys³¹¹ (blue). (D) Plasma-derived HK (light green) or LK (orange; 200 nM) incubated with 40 nM of Plm-Lys³¹¹. (E) Plasma-derived HK (green) or LK (orange; 200 nM) incubated with 40 nM of Plm-Glu³¹¹. (F) Plasma-derived HK (200 nM) incubated with 40 (red), 20 (magenta), or 10 nM (purple) of Plm-Glu³¹¹ or 40 nM of Plg-Lys³¹¹ (blue). (G) Plasma from a patient deficient in HK and LK was supplemented with 600 nM of plasma-derived HK (blue), 2.3 μM of plasma-derived LK (red), HK and LK (green), or vehicle (purple). Plg-Glu³¹¹ or Plg-Lys³¹¹ was added to a final concentration of 600 nM, and tPA (125 nM) was added to activate Plg. In panels A to E, error bars indicate standard errors of the mean for duplicate experiments, each with 2 separate measurements. In panels F and G, error bars indicate standard errors for 2 experiments.